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Objective To evaluate the blocking effect and mechanism of Soybean-derived Bowman-Birk inhibitor (BBI)on LPS-mediated downregulation for tight junction protein(HT-29 cells)in intestinal epithelial cells(IECs). Methods The toxic effect of LPS and BBI on HT-29 cells was detected by CCK8 Kit.HT-29 cells were pretreated by BBI for 6 hours prior to LPS stimulation, the expression of tight junction protein(ZO-1 and Occludin), TLR4, and MyDD8 was detected by the quantitative real-time polymerase chain reaction(PCR)and Western Blot;activation of NF-κB was measured by Western Blot.Results LPS(1 000ng/mL)and BBI(1 000μg/mL)showed no cytotoxicity on HT-29 cells.LPS could significantly upregulate the expression of TLR4 in HT-29 cells, the up-regulation had time-dose effect, and could significantly downregulate the expression of tight junction protein, the down-regulation effect was directly proportional to the concentration of LPS, could activate NF-κB, and had dose effect, effect of LPS on HT-29 cells could be significantly inhibited by BBI.Conclusion By inhibiting the expression of TLR4 and activation of NF-κB in IECs induced by LPS, BBI can significantly block the LPS-mediated inhibitory effect on tight junction protein in intestinal epithelial cells.
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Objective To investigate the inhibitory effect and mechanism of soybean-derived Bowman-Birk inhibi-tor (BBI)on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells.Methods Cytotoxicity effect of LPS and BBI on intestinal epithelial cells was analyzed by MTT assay.Intestinal epithelial cells were pre-treated with BBI,followed by LPS stimulation,expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1)was detected by quantitative real-time polymerase chain reaction;the activation of NF-κB was measured by pNF-κB-luc system and Western Blot.Results The maximum concentration of LPS (10000 ng/mL)and BBI (1000 μg/mL)had no cytotoxicity effect on intestinal epithelial cells.LPS could potently up-regulate the expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1 ),the up-regulation was positively correlated to the concentration of LPS;LPS-induced expression of inflammatory cytokines in intestinal epithelial cells could achieve the highest level,then decreased over time.The up-regulation of LPS on inflammatory cytokines in intestinal epi-thelial cells had dose-time effect;when intestinal epithelial cells were pretreated by BBI for 6 hours,the inhibitory effect of BBI on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells was most obvious, and had dose-time effect.Conclusion BBI can potently inhibit LPS-induced expression of inflammatory cytokines through inhibiting NF-κB in intestinal epithelial cells.
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Objective To investigate the inhibitory effect and mechanism of soybean-derived Bowman-Birk inhibi-tor (BBI)on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells.Methods Cytotoxicity effect of LPS and BBI on intestinal epithelial cells was analyzed by MTT assay.Intestinal epithelial cells were pre-treated with BBI,followed by LPS stimulation,expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1)was detected by quantitative real-time polymerase chain reaction;the activation of NF-κB was measured by pNF-κB-luc system and Western Blot.Results The maximum concentration of LPS (10000 ng/mL)and BBI (1000 μg/mL)had no cytotoxicity effect on intestinal epithelial cells.LPS could potently up-regulate the expression of inflammatory cytokines(TNF-α,IL-1β,IL-8,and MCP-1 ),the up-regulation was positively correlated to the concentration of LPS;LPS-induced expression of inflammatory cytokines in intestinal epithelial cells could achieve the highest level,then decreased over time.The up-regulation of LPS on inflammatory cytokines in intestinal epi-thelial cells had dose-time effect;when intestinal epithelial cells were pretreated by BBI for 6 hours,the inhibitory effect of BBI on LPS-induced expression of inflammatory cytokines in intestinal epithelial cells was most obvious, and had dose-time effect.Conclusion BBI can potently inhibit LPS-induced expression of inflammatory cytokines through inhibiting NF-κB in intestinal epithelial cells.
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Objective: To study the alkaloid compounds in the roots of Cocculus orbiculatus var. mollis. Methods: The compounds were isolated by silica gel chromatography, and their structures were established by spectroscopic methods. Results: A novel cocsolines-type bisbenzylisoquinoline alkaloid named orbiculatinine (1), along with three known compounds (+) 1, 2- dehydroapateline (2), trilobine (3), and isotrilobine (4), were isolated. Conclusion: Compound 1 is a new compound named orbiculatinine. Compounds 2-4 are isolated from this plant for the first time.
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Objective To evaluate the quality of four domestic and three imported fourth-generation HIV diagnostic reagents.Methods The specificity and sensitivity of these assays were analyzed when testing HIV negative samples and HIV-1 RNA positive samples.The relative seroconversion sensitivity index was analyzed when testing BBI seroconversion panels.Results The sensitivity of seven 4th-generation assays were 100％ (95％ CI:99.86％-100％ ),and one sample at the window period of HIV-1 infection were detected as positive.Of the seven assays,one imported assay exhibited the relative largeδ + value (1.0892),and the small δ+ value were found on the remaining six assays (0.0836-0.3003 ).For the samples negative for HIV antibody,varying degrees of false positives were observed on the seven assays ( specificity:97.80％ -99.60％,δ- value:-1.3803 to -0.4778).When testing the BBI seroconversion panels,the relative seroconversion sensitivity index of domestic assays were -0.500-0,however,which of imported assays were -0.600 and -0.700.Conclusion The seven reagents exhibited high sensitivity and specificity.The 4th generation HIV assays can be used as blood screening reagents to find the samples at window period of HIV-1 infection,thus indicating the certain meaning in reducing the transmission risk of HIV-1 for fourth-generation HIV diagnostic reagents.However,the better efficiency to detect HIV-1 early infection was observed on the imported assays than on the domestic assays.
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Objective To investigate the expression of NF-κB/p65 in nasal NK/T cell lymphomas. Methods Immunohistochemis-try and TUNEL were used to study the expression of NF-KB/p65 and cell apoptosis in 23 nasal NK/T cell lymphoma samples and 14 benign lymph node lesions. Results The NF-KB/p65 positive rates were 43.5% (10/23). The expression of NF-κB/p65 was negative correlated with apoptotic index among 23 nasal NK/T cell lymphomas(P <0. 05). The mean survival period in patients expressed NF-KB/p65 was sig-nificantly shorter than that in negative group(P <0. 05). Conclusions Apoptosis inhibited by overexpression of NF-κB/p65 might be in-volved in the development of nasal NK/T cell lymphoma. NF-κB/p65 expression may be an unfavorable prognostic factor of nasal NK/T cell lymphoma.